4. A compound according to claim 3, wherein; A is --C(O)NHOH; Z is phenyl substituted with 0-1 R.sup.b or pyridyl substituted with 0-1 R.sup.b ; U.sup.a is absent or is O; X.sup.a is absent or is CH.sub.2 or CH.sub.2 CH.sub.2 ; Y.sup.a isabsent or is O; provided that U.sup.a --X.sup.a --Y.sup.a forms a spacer of two or more atoms, other than --CH.dbd.CH-- or --C.ident.C--; Z.sup.a is selected from: phenyl substituted with 0-3 R.sup.c, pyridyl substituted with 0-3 R.sup.c, andquinolinyl substituted with 0-3 R.sup.c ; provided that Z, U.sup.a, Y.sup.a, and Z.sup.a do not combine to form a N--N, N--O, O--N, or O--O group; R.sup.a, at each occurrence, is independently selected from: H, CH.sub.3, and CH.sub.2 CH.sub.3 ; R.sup.a1, at each occurrence, is independently selected from: H, CH.sub.3, and CH.sub.2 CH.sub.3 ; R.sup.a2, at each occurrence, is independently selected from: H, CH.sub.3, and CH.sub.2 CH.sub.3 ; R.sup.c, at each occurrence, is independently selectedfrom: C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, OR.sup.a, Cl, F, Br, .dbd.O, NR.sup.a R.sup.a1, CF.sub.3, (CR.sup.a R.sup.a1).sub.n C(O)R.sup.a1, (CR.sup.a R.sup.a1).sub.n C(O)OR.sup.a, (CR.sup.a R.sup.a1).sub.n C(O)NR.sup.a R.sup.a1,(CR.sup.a R.sup.a1).sub.n S(O).sub.p R.sup.a3, and (CR.sup.a R.sup.a1).sub.n SO.sub.2 NR.sup.a R.sup.a1 ; r, at each occurrence, is selected from 0, 1, 2, and 3; and, n, at each occurrence, is selected from 0, 1, 2, and 3.

5. A compound according to claim 1, wherein the compound is selected from the group: N-hydroxy-6-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}acetyl)-5,6,7,8-tetr ahydropyrido[3,4-b]-pyrazine-7-carboxamide; andN-hydroxy-6-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}sulfonyl)-5,6,7,8-te trahydropyrido[3,4-b]pyrazine-7-carboxamide; or a pharmaceutically acceptable salt form thereof.

6. A pharmaceutical composition, comprising: a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound according to claim 1 or a pharmaceutically acceptable salt form thereof.

8. A pharmaceutical composition, comprising: a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound according to claim 2 or a pharmaceutically acceptable salt form thereof.

9. A pharmaceutical composition, comprising: a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound according to claim 3 or a pharmaceutically acceptable salt form thereof.

10. A pharmaceutical composition, comprising: a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound according to claim 4 or a pharmaceutically acceptable salt form thereof.

11. A pharmaceutical composition, comprising: a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound according to claim 5 or a pharmaceutically acceptable salt form thereof.

This invention relates generally to novel bicyclic hydroxamates as inhibitors of matrix metalloproteinases (MMP), TNF-.alpha. converting enzyme (TACE), aggrecanase or a combination thereof, pharmaceutical compositions containing the same, andmethods of using the same.

BACKGROUND OF THE INVENTION

There is now a body of evidence that metalloproteases (MP) are important in the uncontrolled breakdown of connective tissue, including proteoglycan and collagen, leading to resorption of the extracellular matrix. This is a feature of manypathological conditions, such as rheumatoid and osteoarthritis, corneal, epidermal or gastric ulceration; tumor metastasis or invasion; periodontal disease and bone disease. Normally these catabolic enzymes are tightly regulated at the level of theirsynthesis as well as at their level of extracellular activity through the action of specific inhibitors, such as alpha-2-macroglobulins and TIMPs (tissue inhibitors of metalloprotease), which form inactive complexes with the MP's.

Osteo- and Rheumatoid Arthritis (OA and RA respectively) are destructive diseases of articular cartilage characterized by localized erosion of the cartilage surface. Findings have shown that articular cartilage from the femoral heads of patientswith OA, for example, had a reduced incorporation of radiolabeled sulfate over controls, suggesting that there must be an enhanced rate of cartilage degradation in OA (Mankin et al. J. Bone Joint Surg. 1970, 52A, 424-434). There are four classes ofprotein degradative enzymes in mammalian cells: serine, cysteine, aspartic and metalloproteases. The available evidence supports that it is the metalloproteases that are responsible for the degradation of the extracellular matrix of articular cartilagein OA and RA. Increased activities of collagenases and stromelysin have been found in OA cartilage and the activity correlates with severity of the lesion (Mankin et al. Arthritis Rheum. 1978, 21, 761-766, Woessner et al. Arthritis Rheum. 1983, 26,63-68 and Woessner et al. Arthritis Rheum. 1984, 27, 305-312). In addition, aggrecanase has been identified as providing the specific cleavage product of proteoglycan found in RA and OA patients (Lohmander L. S. et al. Arthritis Rheum. 1993, 36,1214-22).

Therefore, metalloproteases (MP) have been implicated as the key enzymes in the destruction of mammalian cartilage and bone. It can be expected that the pathogenesis of such diseases can be modified in a beneficial manner by the administrationof MP inhibitors, and many compounds have been suggested for this purpose (see Wahl et al. Ann. Rep. Med. Chem. 1990, 25, 175-184, AP, San Diego).

Tumor necrosis factor-.alpha. (TNF-.alpha.) is a cell-associated cytokine that is processed from a 26 kd precursor form to a 17 kd active form. TNF-.alpha. has been shown to be a primary mediator in humans and in animals, of inflammation,fever, and acute phase responses, similar to those observed during acute infection and shock. Excess TNF-.alpha. has been shown to be lethal. There is now considerable evidence that blocking the effects of TNF-.alpha. with specific antibodies can bebeneficial in a variety of circumstances including autoimmune diseases such as rheumatoid arthritis (Feldman et al, Lancet 1994, 344, 1105) and non-insulin dependent diabetes melitus. (Lohmander, L. S. et al. Arthritis Rheum. 1993, 36, 1214-22) andCrohn's disease (MacDonald et al. Clin. Exp. Immunol. 1990, 81, 301).

Since excessive TNF-.alpha. production has been noted in several disease conditions also characterized by MMP-mediated tissue degradation, compounds which inhibit both MMPs and TNF-.alpha. production may also have a particular advantage indiseases where both mechanisms are involved.

Prostaglandins (PG) play a major role in the inflammation process and the inhibition of PG production has been a common target of anti-inflammatory drug discovery. Many NSAIDS have been found to prevent the production of PG by inhibiting theenzyme cyclooxygenase (COX). Among the two isoforms of COXs, COX-1 is constitutively expressed. COX-2 is an inducible isozyme associated with inflammation. Selective COX-2 inhibitor was believed to maintain the efficacy of traditional NSAIDs, whichinhibit both isozymes, and produce fewer and less drastic side effects. As a result, development of selective COX-2 inhibitors has attracted major interest in the pharmaceutical industry. Because of the significant roles of PGs and TNF-.alpha. ininflammation, combined use of COX-2 and TACE inhibitors may have superior efficacy to either therapy alone in some inflammatory diseases.

wherein Ar is carbocyclic or heterocyclic aryl; R and R.sub.1 together with the chain to which they are attached form a 1,2,3,4-tetrahydroisoquinoline; R.sub.2 can be hydrogen. The compounds of U.S. Pat. Nos. 5,506,242, 5,552,419, and5,672,615 are not considered to be part of the presently claimed invention.

wherein A can be HO--NH--C(O)--; R.sup.1 can be R.sup.2 --X--Ph--B--; X is a bond or a single chain atom spacer; R.sup.2 is unsubstituted or substituted phenyl; and, Q forms a ring as defined in WO97/18194. These compounds are not considered tobe part of the present invention.

wherein R.sub.1 is hydrogen; R.sub.2 can be hydrogen; W can be arylene or heteroarylene bridge between two adjacent carbons forming a fused ring. These compounds are not considered to be part of the present invention.

DE 19542189 presents matrix metalloproteases inhibitors of the formula: ##STR5##

wherein R.sup.1 can be R.sup.5 --X--Ph--A--; A is C.sub.1-4 alkylene or --CH.dbd.CH--; X is a bond or a single chain atom spacer. These compounds are not considered to be part of the present invention.

wherein m and n are identical or different, represent 0, 1, 2; A represents an aryl ring or heterocycle; X represents --SO.sub.2 --, --CO--, --SO.sub.2 NH--; and R.sub.5 represents optionally substituted alkyl, (C.sub.3-7)cycloalkyl, aryl orheterocyclic. These compounds are not considered to be part of the present invention.

wherein R.sup.1 is unsubstituted or substituted phenyl or hetereocycle; A is a bond, O, --CH.dbd.CH--, or --C.ident.C--; B is C.sub.1-3 alkylene, --O--C.sub.1-5 alkylene- or --CH.dbd.CH--; X is --CH.dbd.CH--, oxygen or sulfur; and R.sup.2 can beHO(H)N--. These compounds are not considered to be part of the present invention.

wherein X can be SO.sub.2 ; Y is aryl or heteroaryl; Z can be O, NH, CH.sub.2 or S; R.sub.2 can be hydrogen; R.sub.4 and R.sub.5 can be hydrogen; and R.sub.1 and R.sub.3 together with the atoms to which they are attached can form a divalentmoiety of the formula: ##STR9##

wherein A is aryl or heteroaryl. These compounds are defined as being useful as TACE and MMP inhibitors. These compounds are not considered to be part of the present invention.

WO00/23443 illustrates TNF-.alpha. production inhibitors of the formula: ##STR10##

wherein R.sup.1 is an optionally substituted C.sub.1-6 alkyl, phenyl, or heteroaryl. These compounds are not considered to be part of the present invention.

wherein R.sub.2 can be --NHOH; and Ar.sub.1 can be phenyl. These compounds are not considered to be part of the present invention.

The compounds of the present invention act as inhibitors of MPs, in particular TACE, MMPs, and/or aggrecanase. These novel molecules are provided as anti-inflammatory compounds and cartilage protecting therapeutics. The inhibition ofaggrecanase, TACE, and other metalloproteases by molecules of the present invention indicates they are anti-inflammatory and should prevent the degradation of cartilage by these enzymes, thereby alleviating the pathological conditions of OA and RA.

The present invention provides pharmaceutical compositions comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of at least one of the compounds of the present invention or a pharmaceutically acceptable salt orprodrug form thereof.

The present invention provides a method for treating inflammatory disorders, comprising: administering to a host, in need of such treatment, a therapeutically effective amount of at least one of the compounds of the present invention or apharmaceutically acceptable salt or prodrug form thereof.

The present invention provides a method of treating a condition or disease mediated by MMPs, TACE, aggrecanase, or a combination thereof in a mammal, comprising: administering to the mammal in need of such treatment a therapeutically effectiveamount of a compound of the present invention or a pharmaceutically acceptable salt or prodrug form thereof.

The present invention provides a method comprising: administering a compound of the present invention or a pharmaceutically acceptable salt or prodrug form thereof in an amount effective to treat a condition or disease mediated by MMPs, TACE,aggrecanase, or a combination thereof.

The present invention provides a method for treating inflammatory disorders, comprising: administering, to a host in need of such treatment, a therapeutically effective amount of one of the compounds of the present invention, in combination withone or more additional anti-inflammatory agents selected from selective COX-2 inhibitors, interleukin-1 antagonists, dihydroorotate synthase inhibitors, p38 MAP kinase inhibitors, TNF-.alpha. inhibitors, TNF-.alpha. sequestration agents, andmethotrexate.

The present invention provides novel compounds of the present invention for use in therapy.

The present invention provides the use of novel compounds of the present invention for the manufacture of a medicament for the treatment of a condition or disease mediated by MMPs, TACE, aggrecanase, or a combination thereof.

These and other objects, which will become apparent during the following detailed description, have been achieved by the inventors' discovery that compounds of formula (I): ##STR13##

(i) when rings B and C form tetrahydroisoquinoline and A is --C(O)NHOH, then R.sup.1 is other than {4-((2-methyl-4-quinolinyl)methoxy)phenyl}acetyl, {(2-hydroxybenzoyl)amino)}benzenesulfonyl, {(4-fluorophenyl)methoxy}benzenesulfonyl, or{(4-methoxyphenyl)carbamate}benzenesulfonyl;

(ii) when rings B and C form tetrahydro-furo[2,3-c]pyridine, A is --C(O)NHOH, and U is SO.sub.2, then Z is other than phenyl;

(iii) when rings B and C form tetrahydro-1H-[1,4]benzodiazepine, A is --C(O)NHOH, U is SO.sub.2, then Z is other than phenyl;

(iv) when U is SO.sub.2, then U.sup.a --X.sup.a --Y.sup.a is other than --OCH.sub.2 --C.ident.C--, --NHCH.sub.2 --C.ident.C--, --CH.sub.2 CH.sub.2 --C.ident.C-- or --SCH.sub.2 --C.ident.C--;

(i) when rings B and C form tetrahydroisoquinoline and A is --C(O)NHOH, then R.sup.1 is other than {4-((2-methyl-4-quinolinyl)methoxy)phenyl}acetyl, {(2-hydroxybenzoyl)amino)}benzenesulfonyl, {(4-fluorophenyl)methoxy}benzenesulfonyl, or{(4-methoxyphenyl)carbamate}benzenesulfonyl;

(ii) when rings B and C form tetrahydro-furo[2,3-c]pyridine, A is --C(O)NHOH, and U is SO.sub.2, then Z is other than phenyl. [3] In another embodiment, the present invention provides a novel compound, wherein; A is selected from: --C(O)NHOH,and --N(OH)CHO; U--X is SO.sub.2, C(O), or C(O)CH.sub.2 ; Z is phenyl substituted with 0-3 R.sup.b or pyridyl substituted with 0-3 R.sup.b ; U.sup.a is absent or is selected from: O, NR.sup.a1, C(O), C(O)NR.sup.a1, S(O).sub.p, and S(O).sub.p NR.sup.a1 ;X.sup.a is absent or is selected from: C.sub.1-4 alkylene, C.sub.2 alkenylene, and C.sub.2 alkynylene; provided that U.sup.a --X.sup.a --Y.sup.a forms a spacer of two or more atoms and is other than --CH.dbd.CH-- or --C.ident.C--; Z.sup.a is a C.sub.5-6carbocycle substituted with 0-3 R.sup.c or a 5-10 membered heteroaryl containing from 1-4 heteroatoms selected from the group consisting of N, O, and S(O).sub.p and substituted with 0-3 R.sup.c ; provided that Z, U.sup.a, Y.sup.a, and Z.sup.a do notcombine to form a N--N, N--O, O--N, O--O, S(O).sub.p --O, O--S(O).sub.p or S(O).sub.p --S(O).sub.p group; R.sup.a, at each occurrence, is independently selected from: H and C.sub.1-4 alkyl; R.sup.a1, at each occurrence, is independently selected from: H,C.sub.1-4 alkyl, phenyl and benzyl; R.sup.b, at each occurrence, is independently selected from: C.sub.1-4 alkyl, OR.sup.a, Cl, F, .dbd.O, NR.sup.a R.sup.a1, C(O)R.sup.a, C(O)OR.sup.a, C(O)NR.sup.a R.sup.a1, S(O).sub.2 NR.sup.a R.sup.a1, S(O).sub.pR.sup.a3, and CF.sub.3 ; R.sup.c, at each occurrence, is independently selected from: C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, OR.sup.a, Cl, F, Br, .dbd.O, NR.sup.a R.sup.a1, CF.sub.3, (CR.sup.a R.sup.a1).sub.n C(O)R.sup.a1, (CR.sup.aR.sup.a1).sub.n C(O)OR.sup.a, (CR.sup.a R.sup.a1).sub.n C(O)NR.sup.a R.sup.a1, (CR.sup.a R.sup.a1).sub.n S(O).sub.p R.sup.a3, (CR.sup.a R.sup.a1).sub.n SO.sub.2 NR.sup.a R.sup.a1, and phenyl; and R.sup.5, at each occurrence, is selected from: C.sub.1-4alkyl substituted with 0-2 R.sup.b, and C.sub.1-4 alkyl substituted with 0-2 R.sup.e ; provided that:

(i) when rings B and C form tetrahydroisoquinoline and A is --C(O)NHOH, then R.sup.1 is other than {4-((2-methyl-4-quinolinyl)methoxy)phenyl}acetyl, {(2-hydroxybenzoyl)amino)}benzenesulfonyl or {(4-fluorophenyl)methoxy}benzenesulfonyl;

(ii) when rings B and C form tetrahydro-furo[2,3-c]pyridine, A is --C(O)NHOH, and U is SO.sub.2, then Z is other than phenyl. [4] In another embodiment, the present invention provides a novel compound, wherein; A is --C(O)NHOH; Z is phenylsubstituted with 0-1 R.sup.b or pyridyl substituted with 0-1 R.sup.b ; U.sup.a is absent or is O; X.sup.a is absent or is CH.sub.2 or CH.sub.2 CH.sub.2 ; Y.sup.a is absent or is O; provided that U.sup.a --X.sup.a --Y.sup.a forms a spacer of two or moreatoms, other than --CH.dbd.CH-- or --C.ident.C--; Z.sup.a is selected from: phenyl substituted with 0-3 R.sup.c, pyridyl substituted with 0-3 R.sup.c, and quinolinyl substituted with 0-3 R.sup.c ; provided that Z, U.sup.a, Y.sup.a, and Z.sup.a do notcombine to form a N--N, N--O, O--N, or O--O group; R.sup.a, at each occurrence, is independently selected from: H, CH.sub.3, and CH.sub.2 CH.sub.3 ; R.sup.a1, at each occurrence, is independently selected from: H, CH.sub.3, and CH.sub.2 CH.sub.3 ;R.sup.a2, at each occurrence, is independently selected from: H, CH.sub.3, and CH.sub.2 CH.sub.3 ; R.sup.c, at each occurrence, is independently selected from: C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, OR.sup.a, Cl, F, Br, .dbd.O, NR.sup.aR.sup.a1, CF.sub.3, (CR.sup.a R.sup.a1).sub.n C(O)R.sup.a1, (CR.sup.a R.sup.a1).sub.n C(O)OR.sup.a, (CR.sup.a R.sup.a1).sub.n C(O)NR.sup.a R.sup.a1, (CR.sup.a R.sup.a1).sub.n S(O).sub.p R.sup.a3, and (CR.sup.a R.sup.a1).sub.n SO.sub.2 NR.sup.a R.sup.a1 ;r, at each occurrence, is selected from 0, 1, 2, and 3; and, n, at each occurrence, is selected from 0, 1, 2, and 3; provided that:

(i) when rings B and C form tetrahydroisoquinoline and A is --C(O)NHOH, then R.sup.1 is other than {4-((2-methyl-4-quinolinyl)methoxy)phenyl}acetyl or {(4-fluorophenyl)methoxy}benzenesulfonyl;

(ii) when rings B and C form tetrahydro-furo[2,3-c]pyridine, A is --C(O)NHOH, and U is SO.sub.2, then Z is other than phenyl. [5] In another embodiment, the present invention provides a novel compound selected from the group:N-hydroxy-6-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}acetyl)-5,6,7,8-tetr ahydro-1,6-naphthyridine-7-carboxamide; N-hydroxy-6-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}sulfonyl)-5,6,7,8-te trahydro-1,6-naphthyridine-7-carboxamide;N-hydroxy-7-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}acetyl)-5,6,7,8-tetr ahydro-1,7-naphthyridine-6-carboxamide; N-hydroxy-7-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}sulfonyl)-5,6,7,8-te trahydro-1,7-naphthyridine-6-carboxamide;N-hydroxy-6-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}acetyl)-5,6,7,8-tetr ahydropyrido[3,4-b]-pyrazine-7-carboxamide; N-hydroxy-6-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}sulfonyl)-5,6,7,8-te trahydropyrido[3,4-b]pyrazine-7-carboxamide;N-hydroxy-5-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}acetyl)-4,5,6,7-tetr ahydro[1,3]oxazolo[5,4-c]pyridine-6-carboxamide; N-hydroxy-5-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}sulfonyl)-4,5,6,7-te trahydro[1,3]oxazolo[5,4-c]pyridine-6-carboxamide;N-hydroxy-5-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}acetyl)-4,5,6,7-tetr ahydro[1,3]thiazolo[4,5-c]pyridine-6-carboxamide; and N-hydroxy-5-({4-[(2-methyl-4-quinolinyl)methoxy]phenyl}sulfonyl)-4,5,6,7-tetrahydro[1,3]thiazolo[4,5-c]pyridine-6-carboxamide; or a pharmaceutically acceptable salt form thereof.

In another embodiment, the present invention provides a novel pharmaceutical composition, comprising: a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of the present invention or a pharmaceuticallyacceptable salt form thereof.

In another embodiment, the present invention provides a novel method for treating an inflammatory disorder, comprising: administering to a patient in need thereof a therapeutically effective amount of a compound of the present invention or apharmaceutically acceptable salt form thereof.

In another embodiment, the present invention provides a novel method of treating a condition or disease mediated by MMPs, TACE, aggrecanase, or a combination thereof in a mammal, comprising: administering to the mammal in need of such treatment atherapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt form thereof.

In another embodiment, the present invention provides a novel method comprising: administering a compound of the present invention or a pharmaceutically acceptable salt form thereof in an amount effective to treat a condition or disease mediatedby MMPs, TACE, aggrecanase, or a combination thereof.

In another embodiment, the present invention provides novel compounds of the present invention for use in therapy.

In another embodiment, the present invention provides the use of novel compounds of the present invention for the manufacture of a medicament for the treatment of a condition or disease mediated by MMPs, TACE, aggrecanase, or a combinationthereof.

In another embodiment, the present invention provides a novel article of manufacture, comprising:

(a) a first container;

(b) a pharmaceutical composition located within the first container, wherein the composition, comprises: a first therapeutic agent, comprising: a compound of the present invention or a pharmaceutically acceptable salt form thereof; and,

(c) a package insert stating that the pharmaceutical composition can be used for the treatment of an inflammatory disorder.

In another embodiment, the present invention provides a novel article of manufacture, comprising:

(a) a first container;

(b) a pharmaceutical composition located within the first container, wherein the composition, comprises: a first therapeutic agent, comprising: a compound of the present invention or a pharmaceutically acceptable salt form thereof; and,

(c) a package insert stating that the pharmaceutical composition can be used in combination with a second therapeutic agent to treat an inflammatory disorder.

In another preferred embodiment, the present invention provides a novel article of manufacture, further comprising:

(d) a second container;

wherein components (a) and (b) are located within the second container and component (c) is located within or outside of the second container.

In another embodiment, the present invention provides a method for treating inflammatory disorders, comprising: administering, to a host in need of such treatment, a therapeutically effective amount of one of the compounds of the presentinvention, in combination with one or more additional anti-inflammatory agents selected from selective COX-2 inhibitors, interleukin-1 antagonists, dihydroorotate synthase inhibitors, p38 MAP kinase inhibitors, TNF-.alpha. inhibitors, TNF-.alpha. sequestration agents, and methotrexate.

This invention also encompasses all combinations of preferred aspects of the invention noted herein. It is understood that any and all embodiments of the present invention may be taken in conjunction with any other embodiment to describeadditional even more preferred embodiments of the present invention. It is also understood that each and every element of any embodiment is intended to be a separate specific embodiment. Furthermore, any elements of an embodiment are meant to becombined with any and all other elements from any of the embodiments to describe additional embodiments.

Definitions

The compounds herein described may have asymmetric centers. Compounds of the present invention containing an asymmetrically substituted atom may be isolated in optically active or racemic forms. It is well known in the art how to prepareoptically active forms, such as by resolution of racemic forms or by synthesis from optically active starting materials. Geometric isomers of double bonds such as olefins and C.dbd.N double bonds can also be present in the compounds described herein,and all such stable isomers are contemplated in the present invention. Cis and trans geometric isomers of the compounds of the present invention are described and may be isolated as a mixture of isomers or as separated isomeric forms. All chiral,diastereomeric, racemic forms and all geometric isomeric forms of a structure are intended, unless the specific stereochemistry or isomeric form is specifically indicated. All processes used to prepare compounds of the present invention andintermediates made therein are considered to be part of the present invention.

Preferably, the molecular weight of compounds of the present invention is less than about 500, 550, 600, 650, 700, 750, 800, 850, or 900 grams per mole. More preferably, the molecular weight is less than about 850 grams per mole. Even morepreferably, the molecular weight is less than about 750 grams per mole. Still more preferably, the molecular weight is less than about 700 grams per mole.

The term "substituted," as used herein, means that any one or more hydrogens on the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valency is not exceeded, and that thesubstitution results in a stable compound. When a substituent is keto (i.e., .dbd.O), then 2 hydrogens on the atom are replaced. Keto substituents are not present on aromatic moieties. When a ring system (e.g., carbocyclic or heterocyclic) is said tobe substituted with a carbonyl group or a double bond, it is intended that the carbonyl group or double bond be part (i.e., within) of the ring.

The term "acylation" as used herein describes the functionalization of a primary or secondary amine by reacting it with an "acylator" to form a stable compound. Examples of acylators include (but are not limited to) an acid chloride, acarboxylic acid anhydride, a sulfonyl chloride, a chloroformate, an isocyanate, an isothiocyanate, etc. the product of which is an amide, a sulfonamide, a carbamate, a urea, and a thiourea respectively.

The present invention is intended to include all isotopes of atoms occurring in the present compounds. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation,isotopes of hydrogen include tritium and deuterium. Isotopes of carbon include C-13 and C-14.

The term "independently selected from", "independently, at each occurrence" or similar language, means that the labeled R substitution group may appear more than once and that each appearance may be a different atom or molecule found in thedefinition of that labeled R substitution group. Thus if the labeled R.sup.a substitution group appear four times in a given permutation of Formula I, then each of those labeled R.sup.a substitution groups may be a different group falling in thedefinition of R.sup.a. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

When any variable (e.g., R.sup.6) occurs more than one time in any constituent or formula for a compound, its definition at each occurrence is independent of its definition at every other occurrence. Thus, for example, if a group is shown to besubstituted with 0-2 R.sup.6, then said group may optionally be substituted with up to two R.sup.6 groups and R.sup.6 at each occurrence is selected independently from the definition of R.sup.6. Also, combinations of substituents and/or variables arepermissible only if such combinations result in stable compounds.

When a bond to a substituent is shown to cross a bond connecting two atoms in a ring, then such substituent may be bonded to any atom on the ring. When a substituent is listed without indicating the atom via which such substituent is bonded tothe rest of the compound of a given formula, then such substituent may be bonded via any atom in such substituent. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

In cases wherein there are amines on the compounds of this invention, these can be converted to amine N-oxides by treatment with MCPBA and or hydrogen peroxides to afford other compounds of this invention. Thus, all shown amines are consideredto cover both the shown amine and its N-oxide (N.fwdarw.O) derivative.

As used herein, "alkyl" or "alkylene" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. C.sub.1-10 alkyl (or alkylene), is intended to include C.sub.1,C.sub.2, C.sub.3, C.sub.4, C.sub.5, C.sub.6, C.sub.7, C.sub.8, C.sub.9, and C.sub.10 alkyl groups. Examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl, and s-pentyl. "Haloalkyl" isintended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, substituted with 1 or more halogen (for example --C.sub.v F.sub.w where v=1 to 3 and w=1 to (2v+1)). Examples ofhaloalkyl include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl. "Alkoxy" represents an alkyl group as defined above with the indicated number of carbon atoms attached through an oxygen bridge. C.sub.1-10 alkoxy, is intended to include C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5, C.sub.6, C.sub.7, C.sub.8, C.sub.9, and C.sub.10 alkoxy groups. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy,s-butoxy, t-butoxy, n-pentoxy, and s-pentoxy. "Cycloalkyl" is intended to include saturated ring groups, such as cyclopropyl, cyclobutyl, or cyclopentyl. C.sub.3-7 cycloalkyl, is intended to include C.sub.3, C.sub.4, C.sub.5, C.sub.6, and C.sub.7cycloalkyl groups. "Alkenyl" or "alkenylene" is intended to include hydrocarbon chains of either a straight or branched configuration and one or more unsaturated carbon-carbon bonds which may occur in any stable point along the chain, such as ethenyland propenyl. C.sub.2-10 alkenyl (or alkenylene), is intended to include C.sub.2, C.sub.3, C.sub.4, C.sub.5, C.sub.6, C.sub.7, C.sub.8, C.sub.9, and C.sub.10 alkenyl groups. "Alkynyl" or "alkynylene" is intended to include hydrocarbon chains of eithera straight or branched configuration and one or more triple carbon-carbon bonds which may occur in any stable point along the chain, such as ethynyl and propynyl. C.sub.2-10 alkynyl (or alkynylene), is intended to include C.sub.2, C.sub.3, C.sub.4,C.sub.5, C.sub.6, C.sub.7, C.sub.8, C.sub.9, and C.sub.10 alkynyl groups.

"Halo" or "halogen" as used herein refers to fluoro, chloro, bromo, and iodo; and "counterion" is used to represent a small, negatively charged species such as chloride, bromide, hydroxide, acetate, and sulfate.

As used herein, "carbocycle" or "carbocyclic residue" is intended to mean any stable 3, 4, 5, 6, or 7-membered monocyclic or bicyclic or 7, 8, 9, 10, 11, 12, or 13-membered bicyclic or tricyclic, any of which may be saturated, partiallyunsaturated, or aromatic. Examples of such carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctane, [4.3.0]bicyclononane, [4.4.0]bicyclodecane,[2.2.2]bicyclooctane, fluorenyl, phenyl, naphthyl, indanyl, adamantyl, and tetrahydronaphthyl.

As used herein, the term "heterocycle" or "heterocyclic group" is intended to mean a stable 5, 6, or 7-membered monocyclic or bicyclic or 7, 8, 9, or 10-membered bicyclic heterocyclic ring which is saturated, partially unsaturated or unsaturated(aromatic), and which consists of carbon atoms and 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of N, O and S and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring. The nitrogen and sulfur heteroatoms may optionally be oxidized. The nitrogen atom may be substituted or unsubstituted (i.e., N or NR wherein R is H or another substituent, if defined). The heterocyclic ring may be attached to its pendant group at anyheteroatom or carbon atom that results in a stable structure. The heterocyclic rings described herein may be substituted on carbon or on a nitrogen atom if the resulting compound is stable. A nitrogen in the heterocycle may optionally be quaternized. It is preferred that when the total number of S and O atoms in the heterocycle exceeds 1, then these heteroatoms are not adjacent to one another. It is preferred that the total number of S and O atoms in the heterocycle is not more than 1. As usedherein, the term "aromatic heterocyclic group" or "heteroaryl" is intended to mean a stable 5, 6, or 7-membered monocyclic or bicyclic or 7, 8, 9, or 10-membered bicyclic heterocyclic aromatic ring which consists of carbon atoms and 1, 2, 3, or 4heteroatoms independently selected from the group consisting of N, O and S. It is to be noted that total number of S and O atoms in the aromatic heterocycle is not more than 1.

The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humanbeings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

As used herein, "pharmaceutically acceptable salts" refer to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are notlimited to, mineral or organic acid salts of basic residues such as amines; and alkali or organic salts of acidic residues such as carboxylic acids. The pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternaryammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic,phosphoric, and nitric; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic,2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, and isethionic.

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound that contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acidor base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile arepreferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, the disclosure of which is hereby incorporated by reference.

Since prodrugs are known to enhance numerous desirable qualities of pharmaceuticals (e.g., solubility, bioavailability, manufacturing, etc.) the compounds of the present invention may be delivered in prodrug form. Thus, the present invention isintended to cover prodrugs of the presently claimed compounds, methods of delivering the same and compositions containing the same. "Prodrugs" are intended to include any covalently bonded carriers which release an active parent drug of the presentinvention in vivo when such prodrug is administered to a mammalian subject. Prodrugs the present invention are prepared by modifying functional groups present in the compound in such a way that the modifications are cleaved, either in routinemanipulation or in vivo, to the parent compound. Prodrugs include compounds of the present invention wherein a hydroxy, amino, or sulfhydryl group is bonded to any group that, when the prodrug of the present invention is administered to a mammaliansubject, it cleaves to form a free hydroxyl, free amino, or free sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of alcohol and amine functional groups in the compounds ofthe present invention.

"Stable compound" and "stable structure" are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.

As used herein, "treating" or "treatment" cover the treatment of a disease-state in a mammal, particularly in a human, and include: (a) preventing the disease-state from occurring in a mammal, in particular, when such mammal is predisposed to thedisease-state but has not yet been diagnosed as having it; (b) inhibiting the disease-state, i.e., arresting it development; and/or (c) relieving the disease-state, i.e., causing regression of the disease state.

"Therapeutically effective amount" is intended to include an amount of a compound of the present invention or an amount of the combination of compounds claimed effective to inhibit a desired metalloprotease in a host. The combination ofcompounds is preferably a synergistic combination. Synergy, as described for example by Chou and Talalay, Adv. Enzyme Regul. 22:27-55 (1984), occurs when the effect (in this case, inhibition of the desired target) of the compounds when administered incombination is greater than the additive effect of the compounds when administered alone as a single agent. In general, a synergistic effect is most clearly demonstrated at suboptimal concentrations of the compounds. Synergy can be in terms of lowercytotoxicity, increased anti-inflammatory effect, or some other beneficial effect of the combination compared with the individual components.

Synthesis

The compounds of the present invention can be prepared in a number of ways well known to one skilled in the art of organic synthesis. The compounds of the present invention can be synthesized using the methods described below, together withsynthetic methods known in the art of synthetic organic chemistry, or variations thereon as appreciated by those skilled in the art. Preferred methods include, but are not limited to, those described below. All references cited herein are herebyincorporated in their entirety herein by reference.

The novel compounds of this invention may be prepared using the reactions and techniques described in this section. The reactions are performed in solvents appropriate to the reagents and materials employed and are suitable for thetransformations being effected. Also, in the description of the synthetic methods described below, it is to be understood that all proposed reaction conditions, including choice of solvent, reaction atmosphere, reaction temperature, duration of theexperiment and work up procedures, are chosen to be the conditions standard for that reaction, which should be readily recognized by one skilled in the art. It is understood by one skilled in the art of organic synthesis that the functionality presenton various portions of the molecule must be compatible with the reagents and reactions proposed. Such restrictions to the substituents that are compatible with the reaction conditions will be readily apparent to one skilled in the art and alternatemethods must then be used.

A variety of compounds of formula (I) wherein A is hydroxamic acid group are prepared from the corresponding esters or carboxylic acids using several routes known in the literature (Scheme 1). The methyl ester of 1 (R.sup.11.dbd.Me) is directlyconverted to hydroxamic acid 2 by treatment with hydroxylamine under basic conditions such as KOH or NaOMe in solvents such as methanol. The methyl ester of 1 (R.sup.11.dbd.Me) can also be converted to O-benzyl protected hydroxamic acid withO-benzylhydroxylamine under similar conditions or using Weinreb's trimethylaluminum conditions (Levin, J. I.; Turos, E.; Weinreb, S. M. Syn. Commun. 1982, 12, 989) or Roskamp's bis[bis(trimethylsilyl)amido]tin reagent (Wang, W.-B.; Roskamp, E. J. J.Org. Chem. 1992, 57, 6101). The benzyl ether is removed by methods well known in the literature such as hydrogenation using palladium on barium sulfate in hydrogen, to give compound 2. Alternatively, 2 can be prepared through the carboxylicintermediate 3. Carboxylic acid 3 is converted to 2 via coupling with hydroxylamine, or its protected equivalents (O-benzylhydroxylamine or O-t-butyldimethylsiloxyamine) followed by deprotection. ##STR15##

Compound 1 can be prepared by coupling the corresponding amine with the carboxylic acid derivative as shown in Scheme 2 to form amides. Coupling with PyBOP and base gives the corresponding amide with unhindered carboxylic acids but moresterically hindered acids require more reactive groups such as the corresponding acid chlorides, acid fluorides, or anhydrides. ##STR16##

A variety of compounds of formula I wherein X--Y--Z--U.sup.a --X.sup.a --Y.sup.a --Z.sup.a is a functionalized phenyl group can be prepared by methods described in Scheme 3. Intermediate 5, available from schemes described previously, isconverted to phenol 6 by hydrogenolysis. Phenol 6 is used as a common intermediate for structure diversification. Reaction of 6 with an electrophile R.sup.c --X provides the alkylated product 7, which alternatively can be synthesized by reacting 6 withR.sup.c --OH under Mitsunobu conditions to produce 7. Compound 6 may also be reacted with acylators such as isocyanates to form carbamates, as depicted in product structure 8. Treatment of compound 6 with triflic anhydride yields an intermediate aryltriflate which can participate in the Suzuki reaction with vinyl boronic acids in the presence of Pd(0) catalysts to form intermediate 9. The olefin in intermediate 9 can be reduced directly to compound 10 or alternatively, oxidized with MCPBA to anintermediate epoxide and rearranged with BF.sub.3 OEt.sub.2 to form ketone 11. Esters 7, 8, 10, and 11 are converted to the hydroxamic acids following the sequences outlined in Scheme 1. ##STR17##

An alternative to the amidation of amine 4 as described previously in Scheme 2 is the acylation of 4 with a variety of other functional groups, some of which are depicted in Scheme 4. Esters 12-14 are then converted to their correspondinghydroxamates using the methods described in Scheme 1. ##STR18##

Although the hydroxamic acid of structure 2 is an important component of MMP inhibition, the heterocycle can be synthesized with non-hydroxamate substitution such as a mercaptomethyl functional group. As shown in Scheme 5, the final compound 18may be derived from an ester 4 via hydrolysis of the ester and selective borane reduction of the carboxylic acid to give compound 16. The primary hydroxyl can be transposed to the primary thiol in a number of ways, shown here is Mitsunobu addition ofthioacetic acid followed by hydrolysis to give compound 18. ##STR19##

A series of compounds containing a quinazoline core stucture were assembled using synthetic Scheme 6. 2,3-dimethylquinazoline 19 is treated with N-chlorosuccinimide and dibenzoylperoxide to chlorinate the benzylic positions. Treatment of 20with diethyl(N-acetylamino)malonate under basic conditions followed by sodium hydride annulates the ring to form 21. Deacetylation and hydrolytic decarboxylation are accomplished using HCl(aq) and the carboxylate is re-esterified using acidic MeOH togive 23. Hydroxamate 24 is formed by functionalizing the amine of compound 23 with acylators using the methods outlined in Schemes 2-4 followed by conversion of the ester to the hydroxamic acid 24 as discussed above. ##STR20##

Other substituted quinoline core structures can be made via substituted heteroaryl and heterocyclic .alpha.-amino acids such as 26 in Scheme 7. Many methods exist for making .alpha.-amino acids (see for instance: Easton, C. J.; Chem. Rev., 1997,97,53-82 and references therein) and shown here is the method of O'Donnell (see J. Org. Chem. 1982, 47, 2663-2666). Alkylation of glycine benzophenone imine with a benzylic electrophile yields compound 26 after deprotection of the imine. Treatment ofthe amino ester with formaldehyde results in Pictet-Spengler cyclization to yield the heterocycle 27 that can be acylated using the methods found in Schemes 2-4 and converted to hydroxamate 28 using the methods in Scheme 1. ##STR21##

In Scheme 8 substituted imidazoles were prepared using the procedure of Neuberger et al (Biochem. J. 1944, 38, 309, 312). Protected histidine derivateives such as 29 are functionalized with a variety of R3 groups and deprotected to givecompound 31. Treatment of the amine with formaldehyde results in Pictet-Spengler type cyclization to yield 32 which is esterified under acidic conditions to give 33. Acylation of the amine (Schemes 2,3,4) followed by conversion to the hydroxamate(Scheme 1) give the final hydroxamic acid 34. ##STR22##

The synthesis of core structure 33 is illustrated in Scheme 9 using the method of Klutchko et al (J. Med. Chem. 1986, 29, 1953). The bis-alkyl bromide 29 can be synthesized using methods known to one skilled in the art of organic synthesis. Esterification of the acid chloride with MeOH gives the methyl ester 30 which can participate in a double SN2 reaction to yield the protected heterocycle 31. Hydrogenolytic removal of the benzyl group provides the basic core structure 32 which isfunctionalized by acylation and subsequent hydroxamate formation using the synthetic methods in Schemes 2-4 and Scheme 1 respectively. ##STR23##

When required, separation of the racemic material can be achieved by HPLC using a chiral column or by a resolution using a resolving agent such as camphonic chloride as in Steven D. Young, et al, Antimicrobial Agents and Chemotheraphy, 1995,2602-2605. A chiral compound of Formula I may also be directly synthesized using a chiral catalyst or a chiral ligand, e.g., Andrew S. Thompson, et al, Tetr. Lett. 1995, 36, 8937-8940. Other features of the invention will become apparent in thecourse of the following descriptions of exemplary embodiments that are given for illustration of the invention and are not intended to be limiting thereof.

EXAMPLES

Abbreviations used in the Examples are defined as follows: "1.times." for once, "2.times." for twice, "3.times." for thrice, ".degree. C." for degrees Celsius, "eq" for equivalent or equivalents, "g" for gram or grams, "mg" for milligram ormilligrams, "mL" for milliliter or milliliters, ".sup.1 H" for proton, "h" for hour or hours, "M" for molar, "min" for minute or minutes, "MHz" for megahertz, "MS" for mass spectroscopy, "NMR" for nuclear magnetic resonance spectroscopy, "HPLC" for highperformance liquid chromatography, "rt" for room temperature, "tlc" for thin layer chromatography, "v/v" for volume to volume ratio. ".alpha.", ".beta.", "D", "L", "R" and "S" are stereochemical designations familiar to those skilled in the art.

To a solution of 2,3-dimethylpyrazine (6.27 g, 57.4 mmol) in 100 mL of carbon tetrachloride was added N-chlorosuccinimide (16.7 g) followed by benzoyl peroxide (278 mg, 2 mol %) and the solution was heated to reflux for 16 h. The reaction wascooled, more benzoyl peroxide (278 mg, 2 mol %) was added and the reaction was refluxed for 5 h. The reaction was monitored by TLC until the disappearance of starting pyrazine. The reaction flask was cooled in an ice bath, the solution filtered, and theeluent was concentrated and chromatographed on SiO.sub.2 using 15% EtOAc in hexane. Obtained 1.14 g of the product (11% yield). MS found: (M+H).sup.+ =178.

To a solution of 1b (1.1 g, 2.36 mmol) in 40 mL THF was added NaH (170 mg, 4.25 mmol, 60% oil dispersion) under N.sub.2 and the solution was stirred for 30 min. The reaction was carefully quenched with water and extracted from sat NaHCO.sub.33.times. with EtOAc. The organic layers were dried with MgSO.sub.4, filtered, concentrated, and chromatographed on SiO.sub.2 using 75% EtOAc in hexane to give 640 mg of 1c as an HCl salt (85% yield). MS found: (M+H).sup.+ =322.

Compound 1c (0.557 mmol) was dissolved in 6 M HCl (3 mL) and refluxed for 1.5 h. The reaction was cooled, concentrated in vacuo, taken up in sat HCl in MeOH (10 mL), and refluxed overnight. The reaction was concentrated in vacuo to give thecrude amino ester HCl salt. The crude material was taken up in 10 mL MeOH and HCl (g) was bubbled through the solution for 30 min. After stirring o/n the reaction was concentrated and dried under vacuum. To this crude amino ester was dissolved in 2 mLDMF followed by the addition of {4-[(2-methyl-4-quinolinyl)methoxy]phenyl}acetic acid (205 mg, 0.67 mmol), diisopropyl-ethylamine (486 .mu.L, 2.79 mmol), and lastly PyBOP (377 mg, 0.72 mmol). After stirring o/n, the reaction was partitioned betweenEtOAc and sat NaHCO.sub.3 and separated. The organic phase was dried over MgSO.sub.4, filtered, concentrated under vacuum, and passed thorough a SiO.sub.2 plug using 10% MeOH in DCM. The residue was purified by HPLC to give 68 mg of 1d as a bis-TFAsalt (17% yield for 3 steps). MS found: (M+H).sup.+ =483.

Table 1 contains representative examples of the present invention. Each entry in the table is intended to be paired with each structural formula at the start of the table. For example, example 1 in Table 1 is intended to be paired with each ofthe following formulae A-U.

The compounds of formula I are expected to possess matrix metalloprotease and/or aggrecanase and/or TNF-.alpha. inhibitory activity. The MMP inhibitory activity of the compounds of the present invention is demonstrated using assays of MMPactivity, for example, using the assay described below for assaying inhibitors of MMP activity. The compounds of the present invention are expected to be bioavailable in vivo as demonstrated, for example, using the ex vivo assay described below. Thecompounds of formula I are expected to have the ability to suppress/inhibit cartilage degradation in vivo, for example, as demonstrated using the animal model of acute cartilage degradation described below.

The compounds provided by this invention should also be useful as standards and reagents in determining the ability of a potential pharmaceutical to inhibit MPs. These would be provided in commercial kits comprising a compound of this invention.

Metalloproteinases have also been implicated in the degradation of basement membranes to allow infiltration of cancer cells into the circulation and subsequent penetration into other tissues leading to tumor metastasis (Stetler-Stevenson, Cancerand Metastasis Reviews, 9, 289-303, 1990). The compounds of the present invention should be useful for the prevention and treatment of invasive tumors by inhibition of this aspect of metastasis.

The compounds of the present invention should also have utility for the prevention and treatment of osteopenia associated with matrix metalloprotease-mediated breakdown of cartilage and bone that occurs in osteoporosis patients.

Some compounds of the present invention have been shown to inhibit TNF production in lipopolysacharride stimulated mice, for example, using the assay for TNF induction in mice and in human whole blood as described below.

Some compounds of the present invention have been shown to inhibit aggrecanase, a key enzyme in cartilage breakdown, as determined by the aggrecanase assay described below.

The compounds of the present invention can be administered alone or in combination with one or more additional anti-inflammatory agents. These agents include, but are not limited to, selective COX-2 inhibitors, interleukin-1 antagonists,dihydroorotate synthase inhibitors, p38 MAP kinase inhibitors, TNF-.alpha. inhibitors, and TNF-.alpha. sequestration agents.

By "administered in combination" or "combination therapy" it is meant that a compound of the present invention and one or more additional therapeutic agents are administered concurrently to the mammal being treated. When administered incombination each component may be administered at the same time or sequentially in any order at different points in time. Thus, each component may be administered separately but sufficiently closely in time so as to provide the desired therapeuticeffect.

TNF-.alpha. sequestration agents that may be used in combination with the compounds of this invention, are TNF-.alpha. binding proteins or anti-TNF-.alpha. antibodies. These agents include, but are not limited to, etanercept (Enbrel.RTM.),infliximab (Remicade.RTM.), adalimumab (D2E7), CDP-571 (Humicade.RTM.), and CDP-870.

Other anti-inflammatory agents that may be used in combination with the compounds of this invention, include, but are not limited to, methotrexate, interleukin-1 antagonists (e.g., anakinra (Kineret.RTM.)), dihydroorotate synthase inhibitors(e.g., leflunomide (Arava.RTM.)), and p38 MAP kinase inhibitors.

Administration of the compounds of the present invention (i.e., a first therapeutic agent) in combination with at least one additional therapeutic agent (i.e., a second therapeutic agent), preferably affords an efficacy advantage over thecompounds and agents alone, preferably while permitting the use of lower doses of each (i.e., a synergistic combination). A lower dosage minimizes the potential of side effects, thereby providing an increased margin of safety. It is preferred that atleast one of the therapeutic agents is administered in a sub-therapeutic dose. It is even more preferred that all of the therapeutic agents be administered in sub-therapeutic doses. Sub-therapeutic is intended to mean an amount of a therapeutic agentthat by itself does not give the desired therapeutic effect for the condition or disease being treated. Synergistic combination is intended to mean that the observed effect of the combination is greater than the sum of the individual agents administeredalone.

A compound is considered to be active if it has an IC.sub.50 or K.sub.i value of less than about 10 .mu.M for the inhibition of a desired MP. Preferred compounds of the present invention have K.sub.i 's or IC.sub.50 's of .ltoreq.1 .mu.M. Morepreferred compounds of the present invention have K.sub.i 's or IC.sub.50 's of .ltoreq.0.1 .mu.M. Even more preferred compounds of the present invention have K.sub.i 's or IC.sub.50 's of .ltoreq.0.01 .mu.M. Still more preferred compounds of thepresent invention have K.sub.i 's or IC.sub.50 's of .ltoreq.0.001 .mu.M.

Aggrecanase Enzymatic Assay

A novel enzymatic assay was developed to detect potential inhibitors of aggrecanase. The assay uses active aggrecanase accumulated in media from stimulated bovine nasal cartilage (BNC) or related cartilage sources and purified cartilage aggrecanmonomer or a fragment thereof as a substrate.

The substrate concentration, amount of aggrecanases time of incubation and amount of product loaded for Western analysis were optimized for use of this assay in screening putative aggrecanase inhibitors. Aggrecanase is generated by stimulationof cartilage slices with interleukin-1 (IL-1), tumor necrosis factor alpha (TNF-.alpha.) or other stimuli. Matrix metalloproteinases (MMPs) are secreted from cartilage in an inactive, zymogen form following stimulation, although active enzymes arepresent within the matrix. We have shown that following depletion of the extracellular aggrecan matrix, active MMPs are released into the culture media (Tortorella, M. D. et al. Trans. Ortho. Res. Soc. 1995, 20, 341). Therefore, in order toaccumulate BNC aggrecanase in culture media, cartilage is first depleted of endogenous aggrecan by stimulation with 500 mg/ml human recombinant IL-.beta. for 6 days with media changes every 2 days. Cartilage is then stimulated for an additional 8 dayswithout media change to allow accumulation of soluble, active aggrecanase in the culture media. In order to decrease the amount of other matrix metalloproteinases released into the media during aggrecanase accumulation, agents which inhibit MMP-1, -2,-3, and -9 biosynthesis are included during stimulation. This BNC conditioned media, containing aggrecanase activity is then used as the source of aggrecanase for the assay. Aggrecanase enzymatic activity is detected by monitoring production ofaggrecan fragments produced exclusively by cleavage at the Glu373-Ala374 bond within the aggrecan core protein by Western analysis using the monoclonal antibody, BC-3 (Hughes, C E, et al., Biochem J 306:799-804, 1995). This antibody recognizes aggrecanfragments with the N-terminus, 374ARGSVIL, generated upon cleavage by aggrecanase. The BC-3 antibody recognizes this neoepitope only when it is at the N-terminus and not when it is present internally within aggrecan fragments or within the aggrecanprotein core. Other proteases produced by cartilage in response to IL-1 do not cleave aggrecan at the Glu373-Ala374 aggrecanase site; therefore, only products produced upon cleavage by aggrecanase are detected. Kinetic studies using this assay yield aKm of 1.5+/-0.35 .mu.M for aggrecanase.

To evaluate inhibition of aggrecanase, compounds are prepared as 10 mM stocks in DMSO, water or other solvents and diluted to appropriate concentrations in water. Drug (50 .mu.L) is added to 50 .mu.L of aggrecanase-containing media and 50 .mu.Lof 2 mg/ml aggrecan substrate and brought to a final volume of 200 .mu.L in 0.2 M Tris, pH 7.6, containing 0.4 M NaCl and 40 mM CaCl.sub.2. The assay is run for 4 hr at 37.degree. C., quenched with 20 mM EDTA and analyzed for aggrecanase-generatedproducts. A sample containing enzyme and substrate without drug is included as a positive control and enzyme incubated in the absence of substrate serves as a measure of background.

Removal of the glycosaminoglycan side chains from aggrecan is necessary for the BC-3 antibody to recognize the ARGSVIL epitope on the core protein. Therefore, for analysis of aggrecan fragments generated by cleavage at the Glu373-Ala374 site,proteoglycans and proteoglycan fragments are enzymatically deglycosylated with chondroitinase ABC (0.1 units/10 .mu.g GAG) for 2 hr at 37.degree. C. and then with keratanase (0.1 units/10 .mu.g GAG) and keratanase II (0.002 units/10 .mu.g GAG) for 2 hrat 37.degree. C. in buffer containing 50 mM sodium acetate, 0.1 M Tris/HCl, pH 6.5. After digestion, aggrecan in the samples is precipitated with 5 volumes of acetone and resuspended in 30 .mu.L of Tris glycine SDS sample buffer (Novex.RTM.) containing2.5% beta mercaptoethanol. Samples are loaded and then separated by SDS-PAGE under reducing conditions with 4-12% gradient gels, transferred to nitrocellulose and immunolocated with 1:500 dilution of antibody BC3. Subsequently, membranes are incubatedwith a 1:5000 dilution of goat anti-mouse IgG alkaline phosphatase second antibody and aggrecan catabolites visualized by incubation with appropriate substrate for 10-30 minutes to achieve optimal color development. Blots are quantitated by scanningdensitometry and inhibition of aggrecanase determined by comparing the amount of product produced in the presence versus absence of compound.

Blood is drawn from normal donors into tubes containing 143 USP units of heparin/10 ml. 225 ul of blood is plated directly into sterile polypropylene tubes. Compounds are diluted in DMSO/serum free media and added to the blood samples so thefinal concentration of compounds are 50, 10, 5, 1, 0.5, 0.1, and 0.01 .mu.M. The final concentration of DMSO does not exceed 0.5%. Compounds are preincubated for 15 minutes before the addition of 100 mg/ml LPS. Plates are incubated for 5 hours in anatmosphere of 5% CO.sub.2 in air. At the end of 5 hours, 750 ul of serum free media is added to each tube and the samples are spun at 1200 RPM for 10 minutes. The supernatant is collected off the top and assayed for TNF-alpha production by a standardsandwich ELISA. The ability of compounds to inhibit TNF-alpha production by 50% compared to DMSO treated cultures is given by the IC.sub.50 value.

TNF Induction in Mice

Test compounds are administered to mice either I.P. or P.O. at time zero. Immediately following compound administration, mice receive an I.P. injection of 20 mg of D-galactosamine plus 10 .mu.g of lipopolysaccharide. One hour later, animalsare anesthetized and bled by cardiac puncture. Blood plasma is evaluated for TNF levels by an ELISA specific for mouse TNF. Administration of representative compounds of the present invention to mice results in a dose-dependent suppression of plasmaTNF levels at one hour in the above assay.

MMP Assays

The enzymatic activities of recombinant MMP-1, 2, 3, 7, 8, 9, 10, 12, 13, 14, 15, and 16 were measured at 25.degree. C. with a fluorometric assay (Copeland, R. A. et al. Bioorganic Med. Chem. Lett. 1995, 5, 1947-1952). Final enzymeconcentrations in the assay were between 0.05 and 10 nM depending on the enzyme and the potency of the inhibitor tested. The permissive peptide substrate, MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH.sub.2, was present at a final concentration of 10 .mu.M in allassays. Initial velocities, in the presence or absence of inhibitor, were measured as slopes of the linear portion of the product progress curves. IC50 values were determined by plotting the inhibitor concentration dependence of the fractional velocityfor each enzyme, and fitting the data by non-linear least squares methods to the standard isotherm equation (Copeland, R. A. Enzymes: A practical Introduction to Structure, Mechanism and Data Analysis, Wiley-VHC, New York, 1996, pp 187-223). All of thecompounds studied here were assumed to act as competitive inhibitors of the enzyme, binding to the active site Zn atom as previously demonstrated by crystallographic studies of MMP-3 complexed with related hydroxamic acids (Rockwell, A. et al. J. Am. Chem. Soc. 1996, 118, 10337-10338). Based on the assumption of competitive inhibition, the IC50 values were converted to Ki values as previously described.

Compounds tested in the above assay are considered to be active if they exhibit a K.sub.i of .ltoreq.10 .mu.M. Preferred compounds of the present invention have K.sub.i 's of .ltoreq.1 .mu.M. More preferred compounds of the present inventionhave K.sub.i 's of .ltoreq.0.1 .mu.M. Even more preferred compounds of the present invention have K.sub.i 's of .ltoreq.0.01 .mu.M. Still more preferred compounds of the present invention have K.sub.i 's of .ltoreq.0.001 .mu.M.

Using the methodology described above, a number of compounds of the present invention were found to exhibit K.sub.i 's of .ltoreq.10 .mu.M, thereby confirming the utility of the compounds of the present invention.

The present invention also encompasses an article of manufacture. As used herein, article of manufacture is intended to include, but not be limited to, kits and packages. The article of manufacture of the present invention, comprises: (a) afirst container; (b) a pharmaceutical composition located within the first container, wherein the composition, comprises: a first therapeutic agent, comprising: a compound of the present invention or a pharmaceutically acceptable salt form thereof; and,(c) a package insert stating that the pharmaceutical composition can be used for the treatment of an inflammatory disorder (as defined previously). In another embodiment, the package insert states that the pharmaceutical composition can be used incombination (as defined previously) with a second therapeutic agent to treat an inflammatory disorder. The article of manufacture can further comprise: (d) a second container, wherein components (a) and (b) are located within the second container andcomponent (c) is located within or outside of the second container. Located within the first and second containers means that the respective container holds the item within its boundaries.

The first container is a receptacle used to hold a pharmaceutical composition. This container can be for manufacturing, storing, shipping, and/or individual/bulk selling. First container is intended to cover a bottle, jar, vial, flask, syringe,tube (e.g., for a cream preparation), or any other container used to manufacture, hold, store, or distribute a pharmaceutical product.

The second container is one used to hold the first container and, optionally, the package insert. Examples of the second container include, but are not limited to, boxes (e.g., cardboard or plastic), crates, cartons, bags (e.g., paper or plasticbags), pouches, and sacks. The package insert can be physically attached to the outside of the first container via tape, glue, staple, or another method of attachment, or it can rest inside the second container without any physical means of attachmentto the first container. Alternatively, the package insert is located on the outside of the second container. When located on the outside of the second container, it is preferable that the package insert is physically attached via tape, glue, staple, oranother method of attachment. Alternatively, it can be adjacent to or touching the outside of the second container without being physically attached.

The package insert is a label, tag, marker, etc. that recites information relating to the pharmaceutical composition located within the first container. The information recited will usually be determined by the regulatory agency governing thearea in which the article of manufacture is to be sold (e.g., the United States Food and Drug Administration). Preferably, the package insert specifically recites the indications for which the pharmaceutical composition has been approved. The packageinsert may be made of any material on which a person can read information contained therein or thereon. Preferably, the package insert is a printable material (e.g., paper, plastic, cardboard, foil, adhesive-backed paper or plastic, etc.) on which thedesired information has been formed (e.g., printed or applied).

Dosage and Formulation

The compounds of the present invention can be administered orally using any pharmaceutically acceptable dosage form known in the art for such administration. The active(ingredient can be supplied in solid dosage forms such as dry powders,granules, tablets or capsules, or in liquid dosage forms, such as syrups or aqueous suspensions. The active ingredient can be administered alone, but is generally administered with a pharmaceutical carrier. A valuable treatise with respect topharmaceutical dosage forms is Remington's Pharmaceutical Sciences, Mack Publishing.

The compounds of the present invention can be administered in such oral dosage forms as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups,and emulsions. Likewise, they may also be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramuscular form, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts. An effective butnon-toxic amount of the compound desired can be employed as an antiinflammatory and antiarthritic agent.

The compounds of this invention can be administered by any means that produces contact of the active agent with the agent's site of action in the body of a mammal. They can be administered by any conventional means available for use inconjunction with pharmaceuticals, either as individual therapeutic agents or in a combination of therapeutic agents. They can be administered alone, but generally administered with a pharmaceutical carrier selected on the basis of the chosen route ofadministration and standard pharmaceutical practice.

The dosage regimen for the compounds of the present invention will, of course, vary depending upon known factors, such as the pharmacodynamic characteristics of the particular agent and its mode and route of administration; the species, age, sex,health, medical condition, and weight of the recipient; the nature and extent of the symptoms; the kind of concurrent treatment; the frequency of treatment; the route of administration, the renal and hepatic function of the patient, and the effectdesired. An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter, or arrest the progress of the condition.

By way of general guidance, the daily oral dosage of each active ingredient, when used for the indicated effects, will range between about 0.001 to 1000 mg/kg of body weight, preferably between about 0.01 to 100 mg/kg of body weight per day, andmost preferably between about 1.0 to 20 mg/kg/day. For a normal male adult human of approximately 70 kg of body weight, this translates into a dosage of 70 to 1400 mg/day. Intravenously, the most preferred doses will range from about 1 to about 10mg/kg/minute during a constant rate infusion. Advantageously, compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three, or four times daily.

The compounds for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches wall known to those of ordinary skill in thatart. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.

In the methods of the present invention, the compounds herein described in detail can form the active ingredient, and are typically administered in admixture with suitable pharmaceutical diluents, excipients, or carriers (collectively referred toherein as carrier materials) suitably selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.

For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose,magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like; for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such asethanol, glycerol, water, and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents, and coloring agents can also be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugarssuch as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate,sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

The compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety ofphospholipids, such as cholesterol, stearylamine, or phosphatidylcholines.

Compounds of the present invention may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamidephenol,or polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore, the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid,polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacylates, and crosslinked or amphipathic block copolymers of hydrogels.

Dosage forms (pharmaceutical compositions) suitable for administration may contain from about 1 milligram to about 100 milligrams of active ingredient per dosage unit. In these pharmaceutical compositions the active ingredient will ordinarily bepresent in an amount of about 0.5-95% by weight based on the total weight of the composition.

The active ingredient can be administered orally in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions. It can also be administered parenterally, in sterile liquiddosage forms.

Gelatin capsules may contain the active ingredient and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets andcapsules can be manufactured as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from theatmosphere, or enteric coated for selective disintegration in the gastrointestinal tract.

Liquid dosage forms for oral administration can contain coloring and flavoring to increase patient acceptance. In general, water, a suitable oil, saline, aqueous dextrose (glucose), and related sugar solutions and glycols such as propyleneglycol or polyethylene glycols are suitable carriers for parenteral solutions. Solutions for parenteral administration preferably contain a water soluble salt of the active ingredient, suitable stabilizing agents, and if necessary, buffer substances. Antioxidizing agents such as sodium bisulfite, sodium sulfite, or ascorbic acid, either alone or combined, are suitable stabilizing agents. Also used are citric acid and its salts and sodium EDTA. In addition, parenteral solutions can containpreservatives, such as benzalkonium chloride, methyl- or propyl-paraben, and chlorobutanol.

Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference text in this field. Useful pharmaceutical dosage-forms for administration of the compounds of this invention canbe illustrated as follows:

Capsules

Capsules are prepared by conventional procedures so that the dosage unit is 500 milligrams of active ingredient, 100 milligrams of cellulose and 10 milligrams of magnesium stearate.

A large number of unit capsules may also prepared by filling standard two-piece hard gelatin capsules each with 100 milligrams of powdered active ingredient, 150 milligrams of lactose, 50 milligrams of cellulose, and 6 milligrams magnesiumstearate.

Xanthan gum is slowly added into distilled water before adding the active ingredient and the rest of the formulation ingredients. The final suspension is passed through a homogenizer to assure the elegance of the final products.

Gelatin is prepared in hot water. The finely pulverized active ingredient is suspended in the gelatin solution and then the rest of the ingredients are mixed in. The suspension is filled into a suitable packaging container and cooled down toform the gel.

Gelcarin.RTM. is dissolved in hot water (around 80.degree. C.) and then the fine-powder active ingredient is suspended in this solution. Sodium saccharin and the rest of the formulation ingredients are added to the suspension while it is stillwarm. The suspension is homogenized and then filled into suitable containers.

All the ingredients are carefully mixed together to make a homogenous paste.

Soft Gelatin Capsules

A mixture of active ingredient in a digestible oil such as soybean oil, cottonseed oil or olive oil is prepared and injected by means of a positive displacement pump into gelatin to form soft gelatin capsules containing 100 milligrams of theactive ingredient. The capsules are washed and dried.

Tablets

Tablets may be prepared by conventional procedures so that the dosage unit is 500 milligrams of active ingredient, 150 milligrams of lactose, 50 milligrams of cellulose and 10 milligrams of magnesium stearate.

A large number of tablets may also be prepared by conventional procedures so that the dosage unit was 100 milligrams of active ingredient, 0.2 milligrams of colloidal silicon dioxide, 5 milligrams of magnesium stearate, 275 milligrams ofmicrocrystalline cellulose, 11 milligrams of starch and 98.8 milligrams of lactose. Appropriate coatings may be applied to increase palatability or delay absorption.

Injectable

A parenteral composition suitable for administration by injection is prepared by stirring 1.5% by weight of active ingredient in 10% by volume propylene glycol and water. The solution is made isotonic with sodium chloride and sterilized.

The compounds of the present invention may be administered in combination with a second therapeutic agent, especially non-steroidal anti-inflammatory drugs (NSAID's). The compound of Formula I and such second therapeutic agent can beadministered separately or as a physical combination in a single dosage unit, in any dosage form and by various routes of administration, as described above.

The compound of Formula I may be formulated together with the second therapeutic agent in a single dosage unit (that is, combined together in one capsule, tablet, powder, or liquid, etc.). When the compound of Formula I and the secondtherapeutic agent are not formulated together in a single dosage unit, the compound of Formula I and the second therapeutic agent may be administered essentially at the same time, or in any order; for example the compound of Formula I may be administeredfirst, followed by administration of the second agent. When not administered at the same time, preferably the administration of the compound of Formula I and the second therapeutic agent occurs less than about one hour apart, more preferably less thanabout 5 to 30 minutes apart.

Preferably the route of administration of the compound of Formula I is oral. Although it is preferable that the compound of Formula I and the second therapeutic agent are both administered by the same route (that is, for example, both orally),if desired, they may each be administered by different routes and in different dosage forms (that is, for example, one component of the combination product may be administered orally, and another component may be administered intravenously).

The dosage of the compound of Formula I when administered alone or in combination with a second therapeutic agent may vary depending upon various factors such as the pharmacodynamic characteristics of the particular agent and its mode and routeof administration, the age, health and weight of the recipient, the nature and extent of the symptoms, the kind of concurrent treatment, the frequency of treatment, and the effect desired, as described above.

Particularly when provided as a single dosage unit, the potential exists for a chemical interaction between the combined active ingredients. For this reason, when the compound of Formula I and a second therapeutic agent are combined in a singledosage unit they are formulated such that although the active ingredients are combined in a single dosage unit, the physical contact between the active ingredients is minimized (that is, reduced). For example, one active ingredient may be entericcoated. By enteric coating one of the active ingredients, it is possible not only to minimize the contact between the combined active ingredients, but also, it is possible to control the release of one of these components in the gastrointestinal tractsuch that one of these components is not released in the stomach but rather is released in the intestines. One of the active ingredients may also be coated with a sustained-release material which effects a sustained-release throughout thegastrointestinal tract and also serves to minimize physical contact between the combined active ingredients. Furthermore, the sustained-released component can be additionally enteric coated such that the release of this component occurs only in theintestine. Still another approach would involve the formulation of a combination product in which the one component is coated with a sustained and/or enteric release polymer, and the other component is also coated with a polymer such as a lowviscositygrade of hydroxypropyl methylcellulose (HPMC) or other appropriate materials as known in the art, in order to further separate the active components. The polymer coating serves to form an additional barrier to interaction with the other component.

These as well as other ways of minimizing contact between the components of combination products of the present invention, whether administered in a single dosage form or administered in separate forms but at the same time by the same manner,will be readily apparent to those skilled in the art, once armed with the present disclosure.

The present invention also includes pharmaceutical kits useful, for example, in the treatment or prevention of osteoarthritis or rheumatoid arthritis, which comprise one or more containers containing a pharmaceutical composition comprising atherapeutically effective amount of a compound of Formula I. Such kits may further include, if desired, one or more of various conventional pharmaceutical kit components, such as, for example, containers with one or more pharmaceutically acceptablecarriers, additional containers, etc., as will be readily apparent to those skilled in the art. Instructions, either as inserts or as labels, indicating quantities of the components to be administered, guidelines for administration, and/or guidelinesfor mixing the components, may also be included in the kit.

In the present disclosure it should be understood that the specified materials and conditions are important in practicing the invention but that unspecified materials and conditions are not excluded so long as they do not prevent the benefits ofthe invention from being realized.

Although this invention has been described with respect to specific embodiments, the details of these embodiments are not to be construed as limitations. Various equivalents, changes and modifications may be made without departing from thespirit and scope of this invention, and it is understood that such equivalent embodiments are part of this invention.